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0 reviewsSUMMARYComprehensive mapping of neuronal connections across entire nervous systems remains a fundamentalchallenge in neuroscience. Here, we introduce labeling individual neurons with chemical dyes andcontrollable sparseness (LINCS), a technology that achieves rapid, ultrabright, and photostable labelingof specific cell types throughout the entire mouse brain and body. LINCS utilizes an engineered, solubility-enhanced biotin ligase for in vivo biotinylation, followed by rapid whole-mount staining with a highaffinity monovalent streptavidin. When integrated with tissue clearing and light-sheet microscopy, thissystem creates an efficient pipeline for profiling long-range neuronal projections across both the centraland peripheral nervous systems. Furthermore, we developed an adeno-associated virus (AAV) strategyemploying Cas9-mediated Cre knockout to achieve stable sparse labeling, permitting the precisemorphological reconstruction of individual neurons at scale. The LINCS toolkit substantially lowers thebarrier to large-scale connectivity mapping and will accelerate the anatomical and functional dissectionof mammalian neural circuits.
